Veterinary World, EISSN: 2231-0916 RESEARCH ARTICLE

Available at www.veterinaryworld.org/Vol.6/Nov-2013/7.pdf Open Access

Protective role of turmeric on histological, ultrastructural and sero-

biochemical changes in cisplatin-induced nephrotoxicity in female rats
B. Ramya, Y. Anjaneyulu, A. Gopala Reddy1, D. Madhuri, M. Lakshman and P. Shivakumar1

Department of Veterinary Pathology, College of Veterinary Science, Sri Venkateswara Veterinary University,
Rajendranagar, Hyderabad, Andhra Pradesh, India; 1. Department of Vetinary Pharmacology & Toxicology, College of
Veterinary Science, Sri Venkateswara Veterinary University, Rajendranagar, Hyderabad, Andhra Pradesh, India
Corresponding author: P. Shivakumar, e mail: drshiva40@gmail.com
Received: 27-06-2013, Revised: 24-08-2013, Accepted: 26-08-2013, Published online: 05-10-2013

doi: 10.14202/vetworld.2013.865-869 How to cite this article: Ramya B, Anjaneyulu Y, Gopala Reddy A, Madhuri D,
Lakshman M and Shivakumar P (2013) Protective role of turmeric on histological, ultrastructural and sero-biochemical
changes in cisplatin-induced nephrotoxicity in female rats, Veterinary World 6(11): 865-869.

Abstract
Aim: Protective role of turmeric was assessed against cisplatin-induced histological and ultra-structural changes in kidney.
Materials and Methods: A total of 48 rats were divided into 4 groups of 12 rats in each. Group 1 was kept as sham control,
group 2 was treated with cisplatin (@ 2 mg/kg b.wt, intraperitoneally on day 1, 7, 14 and 21), group 3 with turmeric (@ 0.05
mg/kg b.wt. p.o. once daily for 28 days) and group 4 with cisplatin + turmeric (as per above schedule). Blood was collected at
fortnight intervals and serum was separated for estimation of kidney biomarkers. Six rats in each group were then euthanized
on day 14 and 28 for histopathology, and tissue parameters were assayed on day 28.
Results: Thibarbituric acid reacting substances (TBARS), protein carbonyls, serum creatinine, blood urea nitrogen (BUN)
were significantly (P < 0.05) increased, while GSH was significantly (P < 0.05) decreased in group 2 as compared to other
groups. Histological sections of kidney from group 2 rats showed marked inter-tubular haemorrhages, congestion, marked
dilation of tubules, and other lesions of pathological significance. Ultrastructural changes of pathological significance were
also recorded in group 2. Kidney sections of group 4 showed nucleus with uniform size with well differentiated nuclear
membrane and nucleolus, and prominent inter tubular and inter cellular junctions.
Conclusion: The study revealed that cisplatin induces nephrotoxicity due to oxidative stress and turmeric was found
beneficial in countering the adverse effects.
Key words: cisplatin, nephrototoxicity, oxidative stress, turmeric.
Introduction the most useful part of the plant for culinary and
Cancer occurance in India is estimated to be medicinal purposes. Turmeric is obtained from dried
around 2.5 million with over eight lakh new cases and rhizomes of C. longa. Biological properties of turmeric
50 thousand deaths occurring each year [1]. includes antibacterial, antiprotozoan, antiviral, hypolipemic,
Chemotherapy and radiotherapy are the most common hypoglycemic, anticoagulant, antioxidant, antitumour
modalities of cancer treatment. Cisplatin is a platinum and anticarcinogenic activities [10]. Turmeric has
containing drug used in the treatment of various types protective role in cisplatin induced hepato-toxicity [11].
of cancers [2]. Cisplatin based combination chemo- The present study was undertaken in female
therapy regimens are currently used as front line Sprague dawley rats to evaluate the effect of cisplatin
therapy in treatment of various solid tumors [3]. It has on kidney biomarkers and histological, ultrastrucural
dose-dependent cytotoxicity on kidney and other alterations, if any, and to assess the protective role of
organs [4, 5]. The adverse drug reactions were due to turmeric in ameliorating the toxic effects.
the increased production of reactive oxygen species Materials and Methods
leading to oxidative stress [6]. Nephroprotective action
Ethical approval: The protocol of the experiment was
of Merremia emarginata in cisplatin toxicity was
reported by Sudhavani et al. [7]. Ulkanet al. [8] approved by the Institutional Animal Ethics Committee
reported protective role of melatonin, potent ani- according to guidelines given by Committee for the
oxidant in cisplatin induced nephrotoxicity. Various Purpose of Control and Supervision of Experiments on
herbs rich in flavonoids and polyphenols have anti- Animals (CPCSEA).
oxidant properties [9]. Amongst the alternative Chemical Reagents: Cisplatin was obtained from VHB
systems of medicine, the use of herbal medicine has Life Sciences Ltd.,Mumbai. Fresh turmeric was
attracted wide attention in India, in view of the procured from farm and was shade dried and made into
availability of medicinal plants in abundance and their powder. Other chemicals and reagents were obtained
easy accessibility. The rhizome of Curcuma longa is from Qualigens Pvt. Ltd., Mumbai, India.
Copyright: The authors. This article is an open access article licensed
under the terms of the Creative Commons Attribution License Animals: Female rats of Sprague dawley (Rattusnorvegicus)
(http://creativecommons.org/licenses/by/2.0) which permits strain 3 months old (weighing 150-200g) were procured
unrestricted use, distribution and reproduction in any medium,
provided the work is properly cited. from NCLAS- NIN, Tarnaka, Andhra Pradesh.
Veterinary World, EISSN: 2231-0916 865
 Available at www.veterinaryworld.org/Vol.6/Nov-2013/7.pdf
Experimental procedure: A total of 48 Sprague dawley
female rats were randomly divided into 4 groups
consisting of 12 rats in each group. All the groups were
maintained as per the following treatment schedule.
Group 1, Sham control were given saline by
intraperitoneal injection weekly once for 4 weeks,
group 2, Cisplatin control (Cisplatin@ 2 mg / kg body
wt. by i.p injection weekly once for 4 weeks), group 3,
turmeric control (turmeric @ 0.05 ppm by oral gavage
needle daily) and group 4 Cisplatin +turmeric
(Cisplatin @ 2 mg/kg body wt. by intraperitoneal
injection weekly once for 4 weeks + turmeric @ 0.05
ppm body wt. by oral gavage needle daily). Turmeric
was administered by dissolving in DMSO.
Statistical analysis: The data were subjected to
statistical analysis by applying one way ANOVA.
Differences between means tested using Duncan's
multiple comparison tests and significance was set at P
< 0.05 [12].
Blood was collected by retro orbital plexus at
fortnight intervals and serum was separated for
estimation of renal biomarkers [13]. Six rats in each
group were euthanized on day 14 and 28 for
histopathology, while tissue parameters were assayed
on day 28. The tissue pieces were stored in 10%
NBFfor histopathological study. The fixed tissues were
processed and stained with Haematoxylin and Eosin (H
& E) [14], stain for histopathology. To study ultra
structural changes, samples were collected in vials and
fixed in 2.5% gluteraldehyde in 0.05 M phosphate
buffer (pH 7.2) for 24 h at 4o C, after the post-fixation,
samples were dehydrated in a series of graded alcohol
and infiltrated and embedded in Araldite 6005 resin.
Ultra thin sections (50-70 nm thickness) were cut with
a glass knife on a Leica Ultra cut UCT-GA-D/E-1/00
ultra microtome and mounted on grids. Then the
sections were stained with saturated aqueous uranyl
acetate and counter stained with 4% lead citrate and
were observed at various magnifications under a
transmission electron microscope (Model: Hitachi, H-
7500) at RUSKA Lab, College of Veterinary Science,
Hyderabad, India.[15].
After sacrifice, kidneys were immediately
excised, rinsed with ice-cold normal saline and stored
at -500C for further homogenization to estimate
thiobarbituric acid reacting substances (TBARS),
protein carbonyls and reduced glutathione (GSH).
Measurement of glutathione (GSH): Kidney GSH
levels were measured as per the procedure described
by[16]. 100 µl of 25% TCA was added to 400 µl of
kidney homogenate, centrifuged and supernatant is
collected to use as sample. To 2.0 ml of 0.6 mM 5-5'
dithiobis-2-nitrobenzoic acid (DTNB) in 0.2 M sodium
phosphate (pH 8), 0.1 ml of sample and 0.9 ml of 0.2 M
phosphate buffer was added and the absorbance was
read at 412 nm against a reagent blank. The standards
(0.05-5 mg/ml) were also treated in the same way.
Veterinary World, EISSN: 2231-0916
Measurement of Thiobarbituric acid reacting substances
(TBARS): Kidney TBARS levels were measured as per
the procedure described by[17]. 500 µl of supernatant
from the homogenate, 1 ml of 10% TCA and 1 ml of
0.67% thiobarbituric acid were taken in a tightly
Stoppard glass tube. The tube was kept in hot water
bath maintained at 1000C for 45 minutes.Remove the
tube from water bath and the contents are centrifuged
after cooling. The supernatant was collected and read
the absorbance at 532 nm against blank. The
concentration of test samples was obtained using molar
extinction coefficient of MDA.
Measurement of protein carbonyls: Kidney protein
carbonyl levels were measured as per the procedure
described by[18]. 1 ml of homogenate containing 1 mg
protein was added to 4 ml of 10 mMdinitrophenyl
hydrazine (DNPH) in 2.5 M HCl. Samples were
vortexed and incubated at room temperature for 60
minutes in dark. Then protein was precipitated by
adding 5 ml of 20% TCA and centrifuged at 3000 rpm
for 10 min, to collect protein precipitate, which was
washed thrice with 4 ml ofethyl acetate: ethanol (1:1)
solution. Final protein precipitate was re-dissolved in 2
ml of 6 M guanidine HCl in 20 mM of potassium
phosphate and kept at 370C for 10 min, and centrifuged
to remove the insoluble substances and the absorbance
was read at 372 nm against 2.5 M HCl blank. Known
concentration of bovine serum albumin (BSA)
dissolved in 6 M guanidine HCl in 20 mM of potassium
phosphate was used as standard.
Results and Discussion
The rats of group 1 did not reveal any abnormal
symptoms throughout the course of the experiment.
Rats of group 2 showed pronounced depression,
inappetance, loss of condition and also revealed
reduction in feed, water consumption and weight gain.
Two to three days after administration of the drug,
marked polyuria and mild diarrhoea were observed in
rats of group 2. Rats of group 4 also showed these
symptoms, though they were mild to moderate when
compared to group 2. Thorough postmortem was
conducted on all sacrificed rats and lesions of
pathological significance were recorded. Reduction in
size of kidney was appreciable in rats of group 2 as
compared to other groups (Fig.1).
The sections of kidney in group 2 on day 14
revealed moderate congestion (Fig.2) and marked inter
tubular hemorrhages (Fig.3). Few sections revealed
marked tubular dilatation (Fig.4). Sections on day 28
revealed moderate hemorrhages with disrupted tubular
epithelium with few tubules showing moderate
degenerative changes (Fig.5). Few glomeruli revealed
disappearance of capillary network while few with
shrunken glomeruli (Fig.6). The sections of kidney in
group 4 on day 14 showed moderate tubular dilatation
with mild degenerative changes (Fig.7) where as
sections on day 28 showed mild dilatation of tubules
(Fig.8). Transmission electron microscopy of kidney in
866
 Available at www.veterinaryworld.org/Vol.6/Nov-2013/7.pdf

Fig. 1 Photograph of rat kidney Fig. 2 Photomicrograph of kidney Fig. 3 Photomicrograph of kidney Fig. 4 Photomicrograph of kidney
showing reduction in size (Group showing moderate congestion showing marked intertubular showing marked tubular dilation
II, day 28). (Group 2, day 14). H&E X100 haemorrhages (Group 2, day 14) (Group 2, day 14). H&E X100
H&E X100

Fig. 5 Photomicrograph of Kidney Fig. 6. Photomicrograph of Kidney Fig. 7. Photomicrograph of Kidney Fig. 8 Photomicrograph of Kidney
showing hemorrhages and showing shrunken glomeruli and showing moderate tubular dilation showing mild dilation of tubules in
disrupted tubular epithelium absence of capillary network in and mild degenerative changes group 4 on day 28. H&E X100
(Group2, day 28). H&E x 100 few. (Group 2, day 28) H&E X 200 (Group 4, day 14). H&E X 100

Fig. 9 TEM of kidney showing
moderate dilation of tubules,
disruption of nuclear chromatin
material (C), perinuclear clumping
of mitochondria (M), margination
of chromatin material and condensed
endoplasmic reticulum(E) (Group
2, day 28). Uranyl acetate lead
citrate X 8950
Fig. 10 TEM of kidney showing
nuclear changes like pyknosis,
dilation of nuclear pores (NP)
condensation, clumping of nuclear
material (C) and cell without
nucleus (WN) (Group 2, day 28).
Uranyl acetate leadcitrate X 3222
Fig. 11 TEM of kidney showing
loss of cell junction, degeneration
of tubular epithelium, disruption
of chromatin (C), elongattion of
mitochondria (M) with electron
dense granular material and
cytoplasmic vacuolation (V)
(Group 2, day 28). Uranyl acetate
lead citrate X 6265
Fig. 12 TEM of Kidney showing
intact intercellular junction (icj),
pyknotic and swollen nucleus (N),
varied shapes and sizes of
mitochondria (M), condensed
rough endoplasmic reticulum
(RER) and cytoplasmic
vacuolation (V) (Group 2, day 28).
Uranyl acetate lead citrate X 3580

group 2 revealed moderate dilatation of tubules, varied
shapes and sizes of mitochondria, disruption of nuclear
chromatin material, perinuclear clumping of
mitochondria, marked cytoplasmic vacuolation,
margination of chromatin material, disrupted and
condensed endoplasmic reticulum (Fig.9). Few
sections showed nuclear changes like pyknosis,
dilatation of nuclear pores, condensation and clumping
of nuclear material (Fig.10), degeneration of tubular
epithelium, disruption of chromatin, elongation of
mitochondria with electron dense granular material
and cytoplasmic vacuolation (Fig.11), intact intercellular
junction, pyknotic and swollen nucleus, varied shapes
of mitochondria,condensed rough endopl-asmic
reticulum and cytoplasmic vacuolation (Fig.12).
Veterinary World, EISSN: 2231-0916
Sections from rats of group 4 showed nucleus with
uniform size, well differentiated nuclear membrane
and nucleolus, intact inner and outer nuclear membrane
(Fig.13). Few sections also revealed regenerating
mitochondria and intact inter-tubular junction (Fig.14)
and few sections with prominent inter tubular and inter
cellular junctions, prominent nucleus, nucleolus and
regenerating mitochondria (Fig.15). Certain others
showed proximal convoluting tubules with
regenerating brush border, swollen and elongated cilia
and regenerating mitochondria (Fig.16).
The concentration of TBARS and protein carbonyls
in rats of group 2 was significantly increased, whereas
GSH concentration was significantly reduced compared
to other groups. This might be due to accumulation of
867
 Available at www.veterinaryworld.org/Vol.6/Nov-2013/7.pdf

Fig. 13 TEM of kidney showing
nucleus (N) with uniform size, well
differrentiated nuclear membrane
and nucleolus (NL), intact inner
and outer nuclear membranes
(NM) (Group 4, day 28). Uranyl
acetate lead citrate X 4475
Fig. 14 TEM of kidney showing
regenenerating mitochondria (M)
and intact intertubular junction
(itj) (Group 4, day 28). Uranyl
acetate lead citrate X 3580
Fig. 15 TEM of kidney showing
prominent inter tubular and inter
cellular junctions (J), prominent
nucleus (N), nucleolus (NL) and
regenerating mitochondria (M)
Uranyl acetate lead citrate X 6265
Fig. 16 TEM of kidney showing
proximal convoluting tubules with
regenerating brush border (B),
swollen, elongated cilia (C) and
regenerating mitochondria (M).
Uranyl acetate lead citrate X 8950

Table-1. Kidney biomarkers of oxidative stress indifferent groups of rats

Group TBARS Protein carbonyls Reduced Glutatione

(n mol MDA/mg protein) (n mol/mg protein) (n mol/mg protein)
1.Sham Control 5.34± 0.29a 3.39± 0.21a 212.31± 7.31a
2.Cisplatin 11.47±0.72b 9.42 ±0.48b 96.70 ±5.36b
3.Turmeric 4.92±0.22a 3.11 ±0.26a 210.66 ±8.40a
4.Cisplatin+Turmeric 7.53±0.43c 6.47± 0.52c 156.61 ±5.97c
Values are Mean + SE (n = 6); One way ANOVA (SPSS Version 15)

Table-2. Serum biomarkers of nephrotoxicity in different groups of rats

Group BUN (mg/dl) Creatinine (mg/dl)

Day 14 Day 28 Day 14 Day 28
a a a
1. Control 14.36 ±0.75 14.26±0.93 1.36 ±0.18 1.31±0.12a
2.Cisplatin 28.49±2.13b 34.05±2.39b 3.63±0.21b 6.57±0.38b
3.Turmeric 13.45±1.16a 13.73±1.26a 1.35±0.16a 1.18±0.17a
4.Cisplatin +Turmeric 20.40±1.00c 24.37±2.15c 2.42±0.23c 4.27±0.29c
Values are Mean + SE (n = 6); One way ANOVA (SPSS Version 15)
Values with different superscripts differ significantly (P<0.05) in the same column

cisplatin in renal epithelial cells' mitochondria, which
further enhanced Reactive Oxygen Species (ROS)
generation by reduced activity of antioxidant enzymes
and by depleting GSH levels. These findings are in
accordance with the results described previously in rats
[19, 20]. Further, it might be due to the oxidation of
mitochondrial proteins and lipids that caused increase
in protein carbonyls and TBARS, which was indicated
by increased levels of renal biomarkers (creatinine and
BUN) in serum. These findings are in agreement
withearlier reports [21].
Histological sections of kidney in group 2 showed
tubular dilatation, shrunken glomerulus, hyaline casts,
inter tubular congestion and other degenerative
changes. Transmission electron microscopy (TEM)
revealed dense chromatin in nucleus and mitochondrial
changes. As kidney being essential organ of
metabolism and elimination of toxic components,
besides rich in poly unsaturated fatty acids that are the
targets for electrophilic molecules, it served as a major
site of attack for cisplatin that gets activated to reactive
platinum complex in vivo. Accumulation of reactive
metabolites of cisplatin induces alterations in cellular
level that might be due to induction of reactive oxygen
species and subsequent oxidative damage. These
findings are in accordance with the earlier findings of
Veterinary World, EISSN: 2231-0916
Prathibha et al. and Chattopadhyay et al.[22, 23].
Conclusion
There was improvement in group 4 (Cisplatin +
turmeric) in all parameters compared to group 2 (Cisplatin
alone), as turmeric had antioxidant properties through
which it combats ROS which was confirmed by the
results of the study.
Authors' contributions
The present article was part of BR's M.V.Sc research
work. YA designed and approved the experimental
protocol. PS drafted the manuscript and AGR critically
reviewed the manuscript and provided laboratory
animal facilities, DM was the minor research
supervisor who made critical suggestions in
conducting the experiment, ML was the incharge of
Ruska Labs where electron microscopy of the samples
was done. All authors read and approved the final
manuscript.
Acknowledgements
The authors are thankful to the Sri Venkateswara
Veterinary University, Tirupati for funding the
research.
868
 Available at www.veterinaryworld.org/Vol.6/Nov-2013/7.pdf
Competing interests
The authors declare that they have no competing interests.
References
1. Nanda Kumar, A. (1990-1996) National Cancer Registry
Program, Indian Council of Medical Research, Consolidated
Report of Population Based Cancer Registries, New Delhi,
India.
2. Stephen, T. (2005) Cisplatin. Chemical and Engineering
News 83: 52.
3. Noori, S. and Mahboob, T. (2010). Protective role of
carnosine pretreatment on cisplatin- induced kidney damage
in rats. Indian Journal of Clinical Biochemistry 25: 86-91.
4. Pratibha, R., Sameer, R., Padmanabh, V., Rataboli.,
Dayanand, A., Bhiwgade. and Chitra, Y. D. (2006) Enzyme
studies of cisplatin induced hepatic tissue damage in rats.
European Journal of Pharmacology 532:290-293.
5. Naqshbandi, A., Khan, M. W., Rizwan, S., Yusufi, A. N. K.
and Khan, F. (2011) Protective role of fish oil in cisplatin
induced liver toxicity.Biology and Medicine 3:86-97.
6. Zafar, A. B. R., David, B. and Pradip, D. (2010) Prevention of
Cisplatin Induced Nephrotoxicity by Ethanolic Extract of
Embelia ribes Fruits and α Tocopherol in Experimental
Animals. Journal of Complementary and Integrative
Medicine 7:53.
7. Sudhavani, V., Chinnikrishnaiah, V., Raghu, Moorthy, V.,
Raghavendra, H.G. and Ranganayakulu, D. (2010)
Merremiaemarginata burmprotects against cisplatin induced
nephrotoxicity rats. Journal of Advances in Drug Research
1: 2230-7761.
8. Ulkan, K., Ertugrul, K., Zeynep, T., Mehmet, T., Ibrahim H.
O., Okkes, Y., Fikrettin, S., and Kazim, S.(2013) Melatonin
suppresses cisplatin-induced nephrotoxicity via activation of
Nrf-2/HO-1 pathway.Nutrition & Metabolism 10:7.
9. Zsuzsanna, K.,Zsengeller,L. E., Dan, B., Bela, H., Partha, M.
B. K., Samir M. P., Ananth. K., Isaac E. S., Pal, P. (2012)
Cisplatin Nephrotoxicity Involves Mitochondrial Injury with
Impaired Tubular Mitochondrial Enzyme Activity. Journal
of Histochemistryand Cytochemistry 60: 521-529.
10. Deeb, D., Xu, Y. X., Jiang, H., Gao, X., Janakiram, N.,
Chapman, R. A. and Gautam, S. C. (2003) Curcumin
enhances TNF-related apoptosis enhancing ligand-induced
apoptosis in LNCaPprostate cancer cells. Molecular Cancer
Therapeutics 2: 95–103.
11. Ramya, B., Anjaneyulu,Y.,Gopalareddy, A.(2013)A study on
********
Veterinary World, EISSN: 2231-0916
cisplatin-induced hepatotoxicity and protectiverole of
turmeric in rats. International Journal of Pharma and Bio
Sciences 4(1): (b) 133 – 143.
12. Snedecor, G. W. and Cochran, W. G. (1994) Statistical
methods, 8th edition, IOWA State University Press, Amer,
IOWA, USA. Pp. 237-252.
13. Satia, N. C., Dawani, R. R. and Goyal, R. K. (1997)Benefical
effects of clonidine in streptozocine induced Diabetes and
Doca-hypersensitive rats. Journal of Pharmacy and
Pharmacology 49:1030-1035.
14. Culling C F A,Handbook of Histopathological Technique,
(Butterworth & Company (Publishers) Limited Landon)
1957. Pp. 1-50.
15. Bozzola, J. J. and Russell, L. D. 1999. Electron Microscopy:
Principles and Techniques for Biologists. 2ndEdn., Jones and
Bartlett, Boston.
16. Moron, M. S., Depierre, J. W. and Mannervik, B. (1979)
Levels of GSH, glutathione reductase and GST in rat lung
and liver. Biochimica et Biophysica Acta. 582: 67-68.
17. Balasubramanian, K. A., Manohar, M. and Mathan, V. I.
(1988)An unidentified inhibitor of lipid peroxidation in
intestinal mucosa. Biochimicaet Biophysica Acta. 962:51-
58.
18. Levine, R. L., Garland, D., Oliver, C. N., Amici, A., Climent,
I., Lenz, A, G., Ahn, B, W., Shaltiel, S. and Stadtman, E. R.
(1990) Determination of carbonyl concentration in oxidative
proteins. Methods in Enzymology. 186: 464-478.
19. Yousef, M. I., Saad, A. A. and El-Shennawy, L. K. (2009)
Role of grape seedextract in by cisplatininduced oxidative
damagein rats. Food and Chemical Toxicology 47: 1176 –
1183.
20. Marie, H. H. and Prasad, D. (2003) Molecular mechanisms
inCisplatin renal-toxicity.Cancer Therapeutics.1: 47–61.
21. Santos, N. A., Catao, C. S., Martins, N. M., Curti, C.,
Bianchi, M.L. and Santos, A.C. (2007)Cisplatin induced
nephrotoxicity is associated with redox state imbalance,
impairedenergy metabolism and apoptosis in rat kidney
mitochondria. Archieves of Toxicology. 81:495-504.
22. Pratibha, R., Dayanand, A., Bhiwgade., Sameer, K.,
Padmanabh, V., Rataboli. And Chitra, Y. D. (2010) Cisplatin
induced histological changes in renal tissue of rat.
Journal of Cell and Animal Biology.4: 108-111.
23. Chattopadhyay, I., Kaushik, B., Bandyopadhyay, U. and
Banerjee, R.K. (2004) Biological effects and medicinal
properties of turmeric and curcumin, Current Science,87 :44-
53.
869